trisodium ({3-[(carboxylatomethyl)amino]propyl}{3-[(carboxylatomethyl)(C16-18, C18 unsat alkyl)amino]propyl}amino)acetate

Endpoint summary

Description of key information

Skin irritation/Corrosion

No skin irritation / corrosion study is available on Sodium tallowamphopolycarboxyglycinate(Amines, N-[3-[(3-aminopropyl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no2060541-47-9). But reliable in-vivo skin corrosion/irritation studies are available on the two substances with the shortest alkyl chain [Sodium cocoamphopolycarboxyglycinate (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no2098351-38-1) and Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts withCAS no2060541-51-5)], as well as the substance with the longest alkyl chain [Sodium oleylamphopolycarboxyglycinate (1,3-Propanediamine, N1-(3-aminopropyl)-N3-[3-[(9Z)-9-octadecen-1-ylamino]propyl]-, N-(carboxymethyl) derivs., sodium saltswith CAS no 2060541-49-1)]. The available in vivo data on this endpoint is consistent within the group, showing no skin irritating properties. Read across within the amphoteric glycinate substance group is considered valid and theSodium tallowamphopolycarboxyglycinate(Amines, N-[3-[(3-aminopropyl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts withCAS no2060541-47-9) is also regarded as not irritating to skin.

 

Eye irritation/corrosion

Sodium tallowamphopolycarboxyglycinate(Amines, N-[3-[(3-aminopropyl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no2060541-47-9) has been tested both in-vivo and in-vitro studies. The available in-vivo study, the key study, was performed in 1985 under GLP.The effects seen, such as redness in conjunctiva and cornea, were reversible and mild, therefore not generating any classification. Also the other in-vivo study available on the Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts withCAS no2060541-51-5) indicated no eye irritation. In the study performed in 1993, a single instillation of the test material into the eye of the rabbit elicited corneal opacificaton, iridial inflammation and well-defined conjunctival irritation. All reactions had resolved 3 or 7 days after instillation. The effects in the in-vitro studies are mild, and do not indicate Cat 1 classification for eye irritation. Currently in-vivo data is required in order to classify eye irritation Cat 2. The weight of evidence from all the available eye irritation data on this group of substances indicates no classification for eye irritation.      

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Justification for type of information:

Read across (Category)

The available data on this endpoint is consistent within the group. Reliable in vivo skin corrosion/irritation studies are available on the two substances with the shortest alkyl chain, as well as the substance with the longest alkyl chain, showing no skin irritating properties. This data is backed up with in vitro studies on the smallest (shortest alky chain, lowest number of amine and carboxymethylated groups) as well as the biggest structure (longest alkyl-unsaturated chain, highest number of amine and carboxymethylated groups) within the amphoteric, glycinate group. Interpolation of the data within the group is therefore possible. Both the in vitro data as well as the in vivo skin irritation tests indicates low skin irritating properties. All the tests are performed on the technical products (a.i. 40 % aqueous solutions). The neat substances may potentially have more irritating properties. This would be in line with the QSAR models estimating the skin irritating properties on the pure substances to be mildly irritating. But as the substances exclusively are produced, handled and marketed as water based solutions, it is not considered relevant to test the neat substances.

Reason / purpose for cross-reference:

read-across: supporting information

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0.53

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 1 day

Irritant / corrosive response data:

Very slight erythema was noted at all test sites one hour after removal of the wrapping and at the 24-hour observation but persisted at only one test site at the 48 and 72-hour observations. Very slight oedema was noted at all test sites one hour after removal of the wrapping but at only one test site at the 24-hour observation. All test sites appeared normal on day seven.

Interpretation of results:

GHS criteria not met

Conclusions:

Two skin irritation / corrosion studies are available on Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1). The key study is an in-vivo study performed in 2016 according to the OECD Guideline 404 and under GLP. It has reliability rating 1 and classifies the substance as not irritating. This is backed up by an in-vitro study from 2016, as well as the available in-vivo and in-vitro data on the other two substances within the group Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) and Sodium oleylamphopolycarboxyglycinate (1,3-Propanediamine, N1-(3-aminopropyl)-N3-[3-[(9Z)-9-octadecen-1-ylamino]propyl]-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-49-1) all showing consistent negative results.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Justification for type of information:

Read across (Category)

The available data on this endpoint is consistent within the group. Reliable in vivo skin corrosion/irritation studies are available on the two substances with the shortest alkyl chain, as well as the substance with the longest alkyl chain, showing no skin irritating properties. This data is backed up with in vitro studies on the smallest (shortest alky chain, lowest number of amine and carboxymethylated groups) as well as the biggest structure (longest alkyl-unsaturated chain, highest number of amine and carboxymethylated groups) within the amphoteric, glycinate group. Interpolation of the data within the group is therefore possible. Both the in vitro data as well as the in vivo skin irritation tests indicates low skin irritating properties. All the tests are performed on the technical products (a.i. 40 % aqueous solutions). The neat substances may potentially have more irritating properties. This would be in line with the QSAR models estimating the skin irritating properties on the pure substances to be mildly irritating. But as the substances exclusively are produced, handled and marketed as water based solutions, it is not considered relevant to test the neat substances.

Reason / purpose for cross-reference:

read-across: supporting information

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 0 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 1 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Neither irritant nor corrosive effects were observed on the intact skin of three male rabbits (strain NZW) after a contact time of 4 hours. Two of the three rabbits showed no effects at all for any of the dermal irritation endpoints evaluated; the third rabbit had very slight Oedema (score 1) after 24 and 48 hours, which was fully reversed after 72 hours.

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Justification for type of information:

Read across (Category)

The available data on this endpoint is consistent within the group. Reliable in vivo skin corrosion/irritation studies are available on the two substances with the shortest alkyl chain, as well as the substance with the longest alkyl chain, showing no skin irritating properties. This data is backed up with in vitro studies on the smallest (shortest alky chain, lowest number of amine and carboxymethylated groups) as well as the biggest structure (longest alkyl-unsaturated chain, highest number of amine and carboxymethylated groups) within the amphoteric, glycinate group. Interpolation of the data within the group is therefore possible. Both the in vitro data as well as the in vivo skin irritation tests indicates low skin irritating properties. All the tests are performed on the technical products (a.i. 40 % aqueous solutions). The neat substances may potentially have more irritating properties. This would be in line with the QSAR models estimating the skin irritating properties on the pure substances to be mildly irritating. But as the substances exclusively are produced, handled and marketed as water based solutions, it is not considered relevant to test the neat substances.

Reason / purpose for cross-reference:

read-across: supporting information

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 6 days

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0.2

Max. score:

4

Reversibility:

fully reversible within: 2 days

Irritant / corrosive response data:

Well defined erythema with very slight oedema was seen in all three animals on Day 1. These reactions gradually ameliorated and had resolved completely by Day 6.

Other effects:

There were no signs of toxicity or ill health in any rabbit during the observation period.

Interpretation of results:

GHS criteria not met

Conclusions:

Two skin irritation / corrosion studies are available on Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1). The key study is an in-vivo study performed in 2016 according to the OECD Guideline 404 and under GLP. It has reliability rating 1 and classifies the substance as not irritating. This is backed up by an in-vitro study from 2016, as well as the available in-vivo and in-vitro data on the other two substances within the group Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) and Sodium oleylamphopolycarboxyglycinate (1,3-Propanediamine, N1-(3-aminopropyl)-N3-[3-[(9Z)-9-octadecen-1-ylamino]propyl]-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-49-1) all showing consistent negative results.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was performed accosring to US federal methods which are comparable to the OECD guideline and the study was performed under GLP. Observations were made for 72 hours although effects were not fully reversed at that time.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

The animals were observed for 72 hours although the effects in one of the animals had not fully reversed at that time.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: recognised breeders
- Age at study initiation: no data
- Weight at study initiation: 2.0-2.3 kg
- Housing: in grid bottom metal cages, not mentioned if the housing was individual
- Diet (e.g. ad libitum): A commercially ( available antibiotic free pelleted rabbit diet (Product Ref. 680, Dalgety-Spillera Limited), ad libitum
- Water (e.g. ad libitum): Mains drinking water via automatic drinking nozzles in each cage were available ad libitum
- Acclimation period: No data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15-20
- Humidity (%): 70-90
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: June 1985

Female animals were used

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated eye of the animals

Amount / concentration applied:

0.1 ml

Duration of treatment / exposure:

A single dose observed for 72 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

6

Details on study design:

REMOVAL OF TEST SUBSTANCE
not done

SCORING SYSTEM:
Scoring system eye irritation

Cornea: degree of density (area most dense taken for reading)
0 No ulceration or opacity
1 Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 Easily discernible translucent areas, details of iris slightly obscured
3 Nacrous areas, no details of iris visible, size of pupil barely discernible
4 Opaque cornea, iris not discernible through the opacity

Area of Cornea Involved
1 One quarter (or less) but not zero
2 Greater than one quarter but less than half
3 Greater than half but less than three quarters
4 Greater than three quarters, up to whole area

Iris
0 Normal
1 Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive)
2 No reaction to light, haemorrhage, gross destruction (any or all of these)

Conjunctivae
Redness (refers to palpebral and bulbar conjunctivae cornea and iris)

0 Blood vessels normal
1 Some blood vessels definitely hyperaemic
(injected)
2 Diffuse, crimson colour, individual vessels not easily discernible
3 Diffuse, beefy red

Chemosis (lids and/or nictitating membranes)
0 No swelling 0
1 Any swelling above normal (includes nictitating membranes)
2 Obvious swelling with partial eversion of lids
3 Swelling with lids about half-closed
4 Swelling with lids more than half-closed

Discharge
0 No discharge
1 Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2 Discharge with moistening of the lids and hairs just adjacent to lids
3 Discharge with moistening of the lids and hairs a considerable area around the eye


TOOL USED TO ASSESS SCORE: Examination was canfined to a macroscopic observation and aids such as a binocular loupe, slit lamp or fluorescsin,staining were not employed.

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0.15

Max. score:

3

Reversibility:

not fully reversible within: 72 hours

Remarks on result:

other: effects were only observed in one out of 6 animals

Irritation parameter:

conjunctivae score

Remarks:

chemosis

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0.55

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Remarks on result:

other: effects were only observed in one out of 6 animals

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0.33

Max. score:

3

Reversibility:

not fully reversible within: 72 hours

Remarks on result:

other: effects were only observed in one out of 6 animals

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0.05

Max. score:

2

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: effects were only observed in one out of 6 animals

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0

Max. score:

4

Irritant / corrosive response data:

24 hours after dosing iris inflammation and severe conjunctival irritation were apparent in one rabbit of the group, no response to treatment being observed in other, animals of the group. Moderate conjunctival irritation was maintained by the one rabbit throughout the study.

Rabbit No and sex	Region of the eye		Hours after instillation			Average 24-48-72 hours
			24	48	72
840-f	Cornea	Degree of opacity	0	0	0	0
		Area	0	0	0	0
	Iris		0	0	0	0
	Conjunctivae	Redness	0	0	0	0
		Chemosis	0	0	0	0
		Discharge	0	0	0	0
842-f	Cornea	Degree of opacity	0	0	0	0
		Area	0	0	0	0
	Iris		0	0	0	0
	Conjunctivae	Redness	0	0	0	0
		Chemosis	0	0	0	0
		Discharge	0	0	0	0
843-f	Cornea	Degree of opacity	0	0	0	0
		Area	0	0	0	0
	Iris		0	0	0	0
	Conjunctivae	Redness	0	0	0	0
		Chemosis	0	0	0	0
		Discharge	0	0	0	0
845-f	Cornea	Degree of opacity	0	0	0	0
		Area	0	0	0	0
	Iris		1	0	0	0.3
	Conjunctivae	Redness	1	1	1	1
		Chemosis	4*	3	3	3.3
		Discharge	2	2	2	2
846-f	Cornea	Degree of opacity	0	0	0	0
		Area	0	0	0	0
	Iris		0	0	0	0
	Conjunctivae	Redness	0	0	0	0
		Chemosis	0	0	0	0
		Discharge	0	0	0	0
847-f	Cornea	Degree of opacity	0	0	0	0
		Area	0	0	0	0
	Iris		0	0	0	0
	Conjunctivae	Redness	0	0	0	0
		Chemosis	0	0	0	0
		Discharge	0	0	0	0

*Blistering of conjunctival-membranes

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

The results of this study indicate that the test material may have little irritant effect in the eye. The composition of the test material is estimated to be 40% active ingredient (incl NaCl) and 60 % water. The active ingredient is Amines, N-[3-[(3-aminopropyl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts (CAS no 2060541-47-9). The response observed appears to have been in an unusually sensitive animal. The animals were only observed for 72 hours and therefore it can not be judges if the effects are reversible within 21 days. However after 72 hours compared with the 24 reading the severity of the effects had decreased some in the affected animal. The active ingredient Amines, N-[3-[(3-aminopropyl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts (CAS no 2060541-47-9) may not be classified as an irritant to the eye according to GHS.

Executive summary:

According to methods similar to OECD and under GLP 0.1 ml aliquots of the test material were instilled into the right eye of each of six rabbits, the other eye remaining untreated as a control. The irritation produced in the treated eye of all rabbits was assessed 24, 48 and 72 hours after dosing. 24 hours after dosing iris inflammation and severe conjunctival irritation were apparent in one rabbit of the group, no response to treatment being observed in other , animals o f the group. Moderate conjunctival irritation was maintained by the one rabbit throughout the study.

The results of this study indicate that the test material may have little irritant effect in the eye. The response observed appears to have

been in an unusually sensitive animal. The animals were only observed for 72 hours and therefore it can not be judges if the effects are reversible within 21 days. However after 72 hours compared with the 24 reading the severity of the effects had decreased some in the affected animal. The active ingredient Amines, N-[3-[(3-aminopropyl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts (CAS no 2060541-47-9) may not be classified as an irritant to the eye according to GHS.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

20 september 1993 - 4 october 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Read-across (Category)

The available in vivo data on this endpoint is consistent within the group. Two in vivo eye irritation studies are available on two of the substances. The effects seen, such as redness in conjunctiva and cornea, were reversible and mild, therefore not generating any classification. Using this data for read across within the group would be extrapolating data to the extremes of the group. In order to support this read across, in vitro eye irritation studies on all the four substances were performed, both in EpiOcular and Isolated Chicken Eye test. Also the in vitro tests results were consistent within the group, indicating a mild to irritating results. There is no possibility to evaluate the reversibility of the effects seen in in vitro studies. The in vitro tests EpiOcular and Isolated chicken eye tests are considered to over-predict the actual eye irritation potential of the substance shown in the rabbit tests. The effects in the in vitro studies are mild, and do not indicate Cat 1 classification for eye irritation. Currently in vivo data is required in order to classify eye irritation Cat 2. The weight of evidence from the available eye irritation data on this group of substances indicates no classification for eye irritation.

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Amount / concentration applied:

0.1 mL

Observation period (in vivo):

24, 48, 72 h

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

fully reversible within: 5 days

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24-48-72 hours

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: 5 days

Interpretation of results:

GHS criteria not met

Conclusions:

A single instillation of the technical product into the eye of the rabbit elicited corneal opacificaton, iridial inflammation and well-defined conjunctival irritation. All reactions had resolved 3 or 7 days after instillation. The composition of the technical product is 40% active ingredient and 60 % water. The observed results do not warrant classification according to GHS.

Executive summary:

A study was performed to assess the eye irritation potential of a technical product to the rabbit. The method followed was that described in EEC Methods for the determination of toxicity, Amex to Directive 92/69/EEC (OJ No. L383A, 29.12.92), Part 3, Method B.5. Acute toxicity (eye irritation). Three rabbits were each administered a single ocular dose of 0.1 ml of the test substance and observed for seven days after instillation. A single instillation of the technical product into the eye of the rabbit elicited corneal opacificaton, iridial inflammation and well-defined conjunctival irritation. All reactions had resolved 3 or 7 days after instillation. The observed results do not warrant classification according to GHS.

Reference 3

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-05-17 to 2016-05-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guidline study

Qualifier:

according to guideline

Guideline:

other: OECD Guideline for the Testing of Chemicals No. 492: (Reconstructed human Cornea-like Epithelium (RhCE) test method)

Qualifier:

according to guideline

Guideline:

other: EURL ECVAM DB-ALM Method Summary No. 164: EpiOcular™ Eye Irritation Test - Summary

Qualifier:

according to guideline

Guideline:

other: EpiOcular™ Eye Irritation Test (OCL-200-EIT) For the prediction of acute ocular irritation of chemicals

GLP compliance:

yes (incl. QA statement)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Germany)

Amount / concentration applied:

50 µL (undiluted)

Duration of treatment / exposure:

incubation: 30 min.

Observation period (in vivo):

post soak: 12 +/- 2 min.
post treatment: 120 min.

Details on study design:

The test was performed on EpiOcular, a reconstituted three-dimensional human corneal epithelium model. Hereby, the test item was applied topically. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 30 min exposure period and 120 min post-treatment period and compared to those of the concurrent negative controls.

Irritation parameter:

other: Mean Relative Tissue Viability [%] from two tissues

Run / experiment:

In-vitro Eye Irritation: Ocular Irritation Assay using the EpiOcular Human Tissue Model

Value:

7.5

Negative controls validity:

valid

Remarks:

Aqua dest.

Positive controls validity:

valid

Remarks:

Methyl acetate

Remarks on result:

positive indication of irritation

The mixture of 50 µL test item per 1 mL MTT medium showed no reduction of MTT as compared to the solvent. The mixture turned to dark red, but not blue/purple. Although the absorption spectra of the mixture showed a peak at 560 nm with an OD of 2.5343, the NSMTT was not determined since the viability of the test item treated tissues (TM) was below 60%.The mixture of 50 µL test item per 1 mL A. dest. and per 2 mL isopropanol showed no colouring as compared to the solvent.Therefore, NSC equaled 0%.

Conclusions:

In this study under the given conditions the test item showed irritant effects. The test item is classified as “irritant“ in accordance with UN GHS “Category 1” or “Category 2”.

Executive summary:

Ocular irritation potential of the test item was predicted from the relative mean tissue viabilities compared to the negative control tissues concurrently treated with Aqua dest.

The test item showed irritant effects. The mean relative tissue viability (% negative control) was ≤ 60% (7.5%).

The controls confirmed the validity of the study. The mean absolute OD₅₇₀of the two negative control tissues was> 0.8 and < 2.5 (2.082). The mean relative tissue viability (% negative control) of the positive control was < 50% (32.5%). The maximum inter tissue difference of replicate tissues of all dose groups was < 20% (1.6 – 9.4%).

This study is classified as acceptable. This study satisfies the requirement for OECD Guideline for the Testing of Chemicals No. 492: (Reconstructed human Cornea-like Epithelium (RhCE) test method).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation / corrosion

The available data on this endpoint is consistent within the group. Reliable in-vivo skin corrosion/irritation studies are available on the two substances with the shortest alkyl chain [Sodium cocoamphopolycarboxyglycinate (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no2098351-38-1) and Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no2060541-51-5)], as well as the substance with the longest alkyl chain [Sodium oleylamphopolycarboxyglycinate (1,3-Propanediamine, N1-(3-aminopropyl)-N3-[3-[(9Z)-9-octadecen-1-ylamino]propyl]-, N-(carboxymethyl) derivs., sodium saltswith CAS no 2060541-49-1)], showing no skin irritating properties. This data is backed up with in-vitro studies on the smallest (shortest alky, lowest number amine and carboxymethylated groups,CAS no2098351-38-1) as well as the biggest structure (longest alkyl-unsaturated, highest number amine and carboxymethylated groups, CAS no 2060541-49-1) within the amphoteric, glycinate group. Interpolation of the data within the group is therefore possible. Both the in-vitro data as well as the in-vivo skin irritation tests indicates low skin irritating properties. All the tests are performed on the technical products (a.i. 40 % aqueous solutions). The neat substances may potentially have more irritating properties. This would be in line with the QSAR models estimating the skin irritating properties on the pure substances to be mildly irritating. But as the substances exclusively are produced, handled and marketed as water based solutions, it is not considered relevant to test the neat substances.

 

Eye irritation / corrosion

Two in-vivo eye irritation studies are available on two of the amphoteric, glycinate substances:Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) and Sodium tallowamphopolycarboxyglycinate (Amines, N-[3-[(3-aminopro yl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-47-9). The effects seen, such as redness in conjunctiva and cornea, were reversible and mild, therefore not generating any classification. Using this data for read across within the group would be extrapolating data to the extremes of the group. In order to support this read across in-vitro eye irritation studies on all the four substances were performed, both in EpiOcular and Isolated Chicken Eye test. Also the in-vitro tests results were consistent within the group, indicating a mild to irritating results. There is no possibility to evaluate the reversibility of the effects seen in in-vitro studies. The in-vitro tests EpiOcular and Isolated chicken eye tests are considered to over-predict the actual eye irritation potential of the substance shown in the rabbit tests. The effects in the in-vitro studies are mild, and do not indicate Cat 1 classification for eye irritation. Currently in-vivo data is required in order to classify eye irritation Cat 2. The weight of evidence from the available eye irritation data on this group of substances indicates no classification for eye irritation.      

Justification for classification or non-classification

Skin

Based on the above mentioned information, Sodium tallowamphopolycarboxyglycinate (Amines, N-[3-[(3-aminopro yl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-47-9)and the other three substances within the amphoteric, glycinate substane group, are not to beclassified according to Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 and is considered “not irritating” to skin.

 

Eye

The conclusion from the available studies is that the substance Sodium tallowamphopolycarboxyglycinate (Amines, N-[3-[(3-aminopro yl)amino]propyl]-N’-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, N-(carboxymethyl) derivs., sodium salts with CAS no 2060541-47-9) and the other three substances within the amphoteric, glycinate substane group, are not to be classified according to Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 and is considered “not irritating” to eye.